Apparatus and method for preparing microparticles

ABSTRACT

Apparatus and method for preparing microparticles. An emulsion is formed by combining two phases in a static mixing assembly. The static mixing assembly preferably includes a preblending static mixer and a manifold. The emulsion flows out of the static mixing assembly into a quench liquid whereby droplets of the emulsion form microparticles. The residence time of the emulsion in the static mixing assembly is controlled to obtain a predetermined particle size distribution of the resulting microparticles.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is related to co-pending application Ser. No.09/223,276 entitled “Apparatus and Method for Preparing MicroparticlesUsing In-Line Solvent Extraction”, filed on even date herewith, theentirety of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to preparation of microparticles. Moreparticularly, the present invention relates to a method and an apparatusfor preparing microparticles having a more controlled and symmetricalparticle size distribution.

2. Related Art

A variety of methods is known by which compounds can be encapsulated inthe form of microparticles. It is particularly advantageous toencapsulate a biologically active or pharmaceutically active agentwithin a biocompatible, biodegradable wall forming material (e.g., apolymer) to provide sustained or delayed release of drugs or otheractive agents. In these methods, the material to be encapsulated (drugsor other active agents) is generally dissolved, dispersed, oremulsified, using stirrers, agitators, or other dynamic mixingtechniques, in a solvent containing the wall forming material. Solventis then removed from the microparticles and thereafter the microparticleproduct is obtained.

Development of a microencapsulation process suitable for commercialscale production typically requires scaling up, by multiple factors, alaboratory scale process and/or a pilot scale process. The scaled-upprocess will almost always require larger piping and higher flow rates,particularly when the scale factor is very large or if it is desired ornecessary to keep process transfer times similar to the smaller scaleprocesses. Scale-up into new, larger equipment is often unpredictableand achieved in large measure through trial and error. However, theeconomic costs of large-scale trial and error experiments can beprohibitive.

One approach to aiding the scale-up process is to use a static mixer toform an emulsion, as disclosed in U.S. Pat. No. 5,654,008. In the methoddisclosed in U.S. Pat. No. 5,654,008, a first phase, comprising theactive agent and the polymer, and a second phase are pumped through astatic mixer into a quench liquid to form microparticles containing theactive agent. The use of a static mixer to form the emulsion tends tomake the scale-up more predictable and reliable than the scale-up ofother dynamic-mixing processes for making microparticles. However,numerous trials and experiments are still required to completely andaccurately scale-up, such as to commercial scale or by a factor of 20 ormore, a process such as the one disclosed in U.S. Pat. No. 5,654,008.

For a commercial scale process, it is particularly important to controlthe distribution of the size of the microparticles to minimize yieldlosses. For example, microparticles, particularly controlled releasemicroparticles containing an active agent or other type of substance tobe released, can range in size of from about 25 μm to about 250 μm indiameter. For a particular commercial product, the useful or desiredmicroparticle size range can be in the range of, for example, 25-150 μm.Even in an efficient commercial production process, some percentage ofthe microparticles produced will be larger than the upper size limit,and some percentage of the microparticles produced will be smaller thanthe lower size limit, resulting in yield losses. Typically, the morenarrow or tighter the desired microparticle size range, the larger theyield losses that result. These yield losses could be avoided orminimized if a more narrow microparticle size distribution could beachieved. Narrowing the microparticle size distribution eliminates orsignificantly reduces the losses resulting from microparticles that falloutside of the desired microparticle size range.

Thus, there is a need in the art for an improved method and apparatusfor preparing microparticles. There is a particular need in the art foran improved process that can control particle size distribution, andachieve a more narrow particle size distribution. There is a furtherneed in the art for an improved process that can be more quickly,reliably, and accurately scaled-up from a laboratory or pilot scale to acommercial scale. The present invention, the description of which isfully set forth below, solves the need in the art for such improvedmethods and apparatus.

SUMMARY OF THE INVENTION

The present invention relates to an apparatus and method for preparingmicroparticles. In one aspect of the invention, a method of preparingmicroparticles is provided. The method comprises:

preparing a first phase, the first phase comprising an active agent anda polymer;

preparing a second phase;

preparing a quench liquid;

pumping the first phase and the second phase through a first staticmixer to form an emulsion; and

flowing the emulsion through a manifold that includes a plurality ofstatic mixers into the quench liquid whereby droplets of the emulsionform microparticles.

In a further aspect of the present invention, another method forpreparing microparticles is provided. The method comprises:

preparing a first phase, the first phase comprising an active agent anda polymer;

preparing a second phase;

preparing a quench liquid;

combining the first phase and the second phase in a first static mixerto form an emulsion, the emulsion forming an outflow of the first staticmixer;

dividing the outflow of the first static mixer to form at least two flowstreams;

flowing each of the at least two flow streams through a separate secondstatic mixer; and

combining the at least two flow streams with the quench liquid wherebydroplets of the emulsion form microparticles.

In a further aspect of the present invention, a method for controllingparticle size distribution of microparticles is provided. The methodcomprises:

preparing a first phase, the first phase comprising an active agent anda polymer;

preparing a second phase;

preparing a quench liquid;

pumping the first phase and the second phase through a static mixingassembly to form an emulsion;

flowing the emulsion into the quench liquid whereby droplets of theemulsion form microparticles; and

adjusting a residence time of the emulsion in the static mixing assemblyto obtain a predetermined particle size distribution of the resultingmicroparticles, wherein the residence time is equal to a length of thestatic mixing assembly divided by an average velocity of the emulsionthrough the static mixing assembly.

In yet a further aspect of the present invention, a microencapsulatedactive agent prepared by a method for preparing microparticles isprovided. Such a method comprises:

preparing a first phase, the first phase comprising an active agent anda polymer;

preparing a second phase;

preparing a quench liquid;

pumping the first phase and the second phase through a first staticmixer to form an emulsion; and

flowing the emulsion through a manifold that includes a plurality ofstatic mixers into the quench liquid whereby droplets of the emulsionform microparticles.

In yet a further aspect of the present invention, a microencapsulatedactive agent prepared by another method for preparing microparticles isprovided. Such a method comprises:

preparing a first phase, the first phase comprising an active agent anda polymer;

preparing a second phase;

preparing a quench liquid;

combining the first phase and the second phase in a first static mixerto form an emulsion, the emulsion forming an outflow of the first staticmixer;

dividing the outflow of the first static mixer to form at least twosubstantially identical flow streams;

flowing each of the at least two substantially identical flow streamsthrough a separate second static mixer; and

combining the least two substantially identical flow streams with thequench liquid whereby droplets of the emulsion form microparticles.

In still a further aspect of the present invention, a system forpreparing microparticles is provided. The system includes a first andsecond pump, and a first static mixer in fluid communication with eachof the pumps. One of the pumps is configured to pump an organic phaseinto the first static mixer. One of the pumps is configured to pump acontinuous phase into the first static mixer. A manifold, comprising aplurality of static mixers, is in fluid communication with the firststatic mixer. An extraction vessel is in fluid communication with themanifold. The outflow of the first static mixer flows through themanifold into the extraction vessel. The plurality of static mixers inthe manifold can be configured in parallel or in series.

Features and Advantages

It is a feature of the present invention that it can be used to preparemicroparticles, including microparticles containing an active agent.

A significant advantage of the present invention is that it provides amethod for controlling particle size distribution. By controllingparticle size distribution, yield losses, resulting from microparticlesthat fall outside of a desired microparticle size range, can besubstantially reduced or eliminated. This makes the present inventionparticularly useful for commercial products.

The present invention also advantageously allows for use of a morenarrow or tighter target microparticle size range than in conventionalprocesses. Narrowing the limit for the microparticle size rangetypically results in larger yield losses. These yield losses can beavoided or minimized by achieving a more narrow microparticle sizedistribution through the process of the present invention.

The present invention provides a method and apparatus that areparticularly advantageous for scale-up. The parallel path manifold ofthe present invention allows for capacity increases from an established(single path) system without full-scale trial and error experiments innew and different equipment. The total flow rate can be increased fromthe single path system based upon the number of flow streams in themanifold.

BRIEF DESCRIPTION OF THE FIGURES

The present invention is described with reference to the accompanyingdrawings. In the drawings, like reference numbers indicate identical orfunctionally similar elements.

FIG. 1 illustrates flow through a static mixer;

FIG. 2 shows a static mixer suitable for use with the present invention;

FIGS. 3A-3C show various types or designs of static mixing elements;

FIG. 4 depicts a graph of mass median diameter (microns) ofmicroparticle size distribution as a function of average emulsionvelocity (cm/min);

FIG. 5 depicts a graph of width of microparticle size distribution, asexpressed by CoV_(m), for various static mixers as a function ofemulsion residence time (sec) in the static mixer;

FIG. 6 depicts a graph of skewness of microparticle size distributionfor various static mixers as a function of emulsion residence time (sec)in the static mixer; and

FIG. 7 shows an exemplary embodiment of an equipment configuration forpreparing microparticles in accordance with the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Overview

The present invention provides an improved method and apparatus forpreparing microparticles. Maximum yield within a desired microparticlesize range is an important aspect of any process or method for preparingmicroparticles. The size of the resulting microparticles is primarilycontrolled during the emulsification step of the process of the presentinvention. As will be explained in more detail below, the emulsificationstep uses an in-line motionless or static mixer to create an emulsionfrom a first phase, comprising a polymer and a drug or other activeagent, and a second phase, preferably an aqueous solution. The inventorsof the present invention have unexpectedly found that the most dominantinfluential characteristic controlling the dispersity of microparticlesize is the residence time of the emulsion in the static mixer beforethe emulsion is introduced to a quench or solvent-extraction liquid.Increased residence time in the emulsion-forming static mixer decreasespolydispersity, and results in a more symmetrical particle sizedistribution.

The methods of the present invention use a static mixer to combine afirst phase, comprising an active agent and a polymer, with a secondphase to form an emulsion. The static mixer in which the first andsecond phases are combined to form the emulsion may be referred toherein as a “preblending static mixer”. A process for forming anemulsion using a static mixer is described, for example, in U.S. Pat.No. 5,654,008, the entirety of which is incorporated herein byreference. The phase comprising the active agent and the polymer may bereferred to herein as the “organic phase”. The other phase may bereferred to herein as the “continuous phase”.

The emulsion flowing out of the preblending static mixer flows through amanifold containing a plurality of static mixers into a quench liquidwhereby droplets of the emulsion form microparticles. Alternatively, theoutflow of the preblending static mixer is divided to form at least twoflow streams. Each of the flow streams then flows through anotherseparate static mixer. The flow streams are recombined downstream, andcombined with the quench liquid whereby droplets of the emulsion formmicroparticles.

In a particularly preferred embodiment, the diameter of the preblendingstatic mixer is greater than the diameter of the static mixers in themanifold, i.e., greater than the diameter of the static mixers throughwhich the divided flow streams flow. When using a static mixer to forman emulsion, a variety of factors determine emulsion droplet size.Emulsion droplet size determines the size of the resultingmicroparticles. These factors include the density and viscosity of thevarious solutions or phases to be mixed, volume ratio of the phases,interfacial tension between the phases, static mixer parameters (conduitdiameter; length of mixing elements; number of mixing elements), andfluid velocity through the static mixer. The primary controllingvariable is fluid velocity. Droplet size decreases as fluid velocityincreases. Similarly, droplet size increases as fluid velocity (andpressure drop) decreases. Accordingly, the preblending static mixer ispreferably larger in diameter than the static mixers in the manifold inorder to handle the total flow from the feed streams (organic phase andcontinuous phase) at a lower velocity than the smaller diameter staticmixers in the manifold.

In the present invention, a preblending static mixer is used to combinethe organic phase and the aqueous phase to form the emulsion. In oneembodiment, the emulsion is divided into a plurality of flow streams forflow through the manifold. The use of the preblending static mixer priorto the manifold is particularly advantageous because the organic phaseand the aqueous phase are not immediately miscible or homogeneous,making the division of the combined flow stream problematic. Formultiphase (e.g., oil/water) streams, the use of the manifold withoutthe preblending static mixer could result in different compositions ineach static mixer in the manifold. Because the combined organic andaqueous phases is not homogeneous, it would not divide evenly inconventional piping.

The manifold configuration of the present invention is particularlyadvantageous for scale-up. The parallel path manifold of the smallerdiameter static mixers allows for capacity increases from an established(single path) system without full-scale trial and error experiments innew and different equipment. The total flow rate can be increased fromthe single path system based upon the number of flow streams in themanifold.

The present invention advantageously provides a method for controllingparticle size distribution of microparticles. A first phase, comprisingan active agent and a polymer, and a second phase are pumped through astatic mixing assembly to form an emulsion. The emulsion flows into aquench liquid, whereby droplets of the emulsion form microparticles. Byadjusting a residence time of the emulsion in the static mixingassembly, a predetermined particle size distribution of the resultingmicroparticles can be obtained. The residence time is equal to a lengthof the static mixing assembly divided by an average velocity of theemulsion through the static mixing assembly. The particle sizedistribution can be narrowed by increasing the residence time. Theparticle size distribution can be broadened by decreasing the residencetime. The residence time is preferably adjusted by changing the lengthof the static mixing assembly.

To ensure clarity of the description that follows, the followingdefinitions are provided. By “microparticles” or “microspheres” is meantsolid particles that contain an active agent or other substancedispersed or dissolved within a polymer that serves as a matrix orbinder of the particle. The polymer is preferably biodegradable andbiocompatible. By “biodegradable” is meant a material that shoulddegrade by bodily processes to products readily disposable by the bodyand should not accumulate in the body. The products of thebiodegradation should also be biocompatible with the body. By“biocompatible” is meant not toxic to the body, is pharmaceuticallyacceptable, is not carcinogenic, and does not significantly induceinflammation in body tissues. As used herein, “body” preferably refersto the human body, but it should be understood that body can also referto a non-human animal body. By “weight %” or “% by weight” is meantparts by weight per total weight of microparticle. For example, 10 wt. %active agent would mean 10 parts active agent by weight and 90 partspolymer by weight. Unless otherwise indicated to the contrary,percentages (%) reported herein are by weight. By “controlled releasemicroparticle” or “sustained release microparticle” is meant amicroparticle from which an active agent or other type of substance isreleased as a function of time. By “mass median diameter” is meant thediameter at which half of the distribution (volume percent) has a largerdiameter and half has a smaller diameter.

METHOD AND EXAMPLES

The following examples are provided to explain the invention, and todescribe the materials and methods used in carrying out the invention.The examples are not intended to limit the invention in any manner.

Example 1 Static Mixer Tests

A test program was conducted using a variety of static mixers. A staticor motionless mixer consists of a conduit or tube in which is received anumber of static mixing elements. Static mixers provide uniform mixingin a relatively short length of conduit, and in a relatively shortperiod of time. With static mixers, the fluid moves through the mixer,rather than some part of the mixer, such as a blade, moving through thefluid. Flow through one type of static mixer is illustrated in FIG. 1. Apump (not shown) introduces a stream of one or more fluids into a staticmixer 10, as shown generally at 1. The stream is split and forced toopposite outside walls, as shown generally at 2. A vortex is createdaxial to the centerline of static mixer 10, as shown generally at 3. Thevortex is sheared and the process recurs, but with the oppositerotation, as shown generally at 4. The clockwise/counterclockwise motionensures a homogeneous product.

One example of a static mixer is shown in FIG. 2. Static mixer 10includes a number of stationary or static mixing elements 14 arranged ina series within a conduit or pipe 12. The number of static mixingelements can range from 4 to 32 or more. Conduit 12 is circular incross-section and open at opposite ends 18 and 20 for introducing andwithdrawing fluids. Mixing element 14 comprises segments 142. Eachsegment 142 consists of a plurality of generally flat plates or vanes144. The two substantially identical segments 142 are preferably axiallystaggered with respect to each other. A static mixer as shown in FIG. 2is more fully described in U.S. Pat. No. 4,511,258, the entirety ofwhich is incorporated herein by reference.

Each of the static mixers tested displayed characteristic differences indesign, length, diameter, and number of elements per length. A keyaspect to the design of a static mixer is the geometry of the staticmixing elements. The eight static mixers tested used three differenttypes of static mixing elements: scissors elements; helical elements;and layered elements. A scissors mixing element, shown in FIG. 3A, hastwo rows of baffles affixed perpendicular to each other, creating theimpression of a pair of opened scissors. A helical mixing element, shownin FIG. 3B, uses a 180° helical plane with a 90° offset and alternatingdirection of sequential elements. A layered element, shown in FIG. 3C,uses corrugated sheets welded together to form open channels.

The characteristics of the eight static mixers tested are shown below inTable 1. The mixers ranged in size from 2½ inches in length with 5mixing elements to 9¾ inches in length with 24 mixing elements. Byarranging identical static mixers in series, the length and the numberof mixing elements was increased; however, the number of mixing elementsper unit length remained constant. The diameter of the static mixerstested ranged from ¼ inch to ½ inch.

TABLE 1 Characteristics of the Test Mixers # of Mix- Dia- Ele- ElementManu- er # meter Length ments Design Material facturer 1 ¼″ 6″ 24Helical Plastic Cole Palmer 2 ¼″ 9¾″ 24 Relical Stainless TAH Steel 3 ⅜″4¾″ 12 Helical Plastic Cole Palmer 4 ⅜″ 9″ 12 Helical Stainless KenicsSteel 5 ⅜″ 2½″ 5 Layered Stainless Koch Steel 6 ½″ 5″ 12 Helical PlasticTAH 7 ½″ 6″ 12 Helical Plastic Cole Palmer 8 {fraction (5/16)}″ 2⅞″ 12Scissors Plastic Omega

One-hundred gram batches of 40% theoretically loaded risperidonemicroparticles were used to test the static mixers. A 16.7% polymersolution was prepared by dissolving 60 grams of MEDISORB® 7525 DLpolymer (Alkermes, Inc., Blue Ash, Ohio) in ethyl acetate. A 28.6% drugsolution was prepared by dissolving 40 grams of risperidone base(Janssen Pharmaceutica, Beerse, Belgium) in benzyl alcohol. The organicphase was prepared by mixing the drug solution into the polymersolution. The continuous or aqueous phase was a 1% polyvinyl alcohol(PVA) solution containing 6.5% ethyl acetate. The temperature of theorganic and aqueous phases was 25° C. The ratio of the aqueous phase tothe organic phase was 5:1. The quench liquid or extraction solution wasan aqueous solution containing 2.5% ethyl acetate at 25° C. The ratio ofthe volume of extraction solution to batch size was 0.25 liters/gram.

The 100-gram batches were produced by forming an oil-in-water emulsion,and then extracting the solvent from the emulsion droplet to form ahardened polymer/drug microparticle. Two calibrated micropumps fed thestatic mixer under test, one micropump for the organic phase and onemicropump for the aqueous phase. The micropumps were calibrated bymonitoring total volume pumped over a timed period prior to the batchbeing run. A “Y” connection upstream of the static mixer under test wasused to introduce the organic phase into the aqueous phase.

The static mixer under test discharged the resulting emulsion directlyinto an agitated extraction solution. The emulsion remained in theextraction solution for fifteen minutes to harden into microparticles. Asample of the resulting microparticles was tested using a Coulter LS230Small Module particle size analyzer to determine particle sizedistribution. The flow rates were adjusted to target a mass mediandiameter of between about 30 μm and 90 μm. The experimental results forthe eight static mixers tested is shown below in Table 2.

TABLE 2 Summary of Experimental Results Process Conditions Cal. ParticleData Total Cal. Residence @ @ Mixer Diameter Elements Length FlowVelocity Time Median 84% 16% Mean Skew- Number Geometry (in) (#) (in)(ml/min) (cm/min) (sec) (um) (um) (um) CoV_(m) (um) CoV_(x) ness 1Helical 0.250 24 6.00 971 3068 0.298 24.60 43.88 6.07 1.54 25.45 0.6660.259 1 Helical 0.250 24 6.00 464 1467 0.623 54.56 92.01 18.83 1.3455.85 0.606 0.260 1 Helical 0.250 24 6.00 329 1039 0.880 87.76 159.1033.30 1.43 97.33 0.667 0.846 2 Helical 0.250 24 9.75 960 3033 0.49057.28 120.80 16.65 1.82 67.37 0.748 0.751 3 Helical 0.375 12 4.75 11401600 0.452 43.20 88.64 13.59 1.74 50.01 0.723 0.737 3 Helical 0.375 124.75 1213 1703 0.425 47.93 89.66 18.40 1.49 53.2 0.627 0.590 3 Helical0.375 12 4.75 1215 1706 0.424 47.13 90.58 15.14 1.60 52.11 0.677 0.532 3Helical 0.375 60 23.50 1133 1591 2.251 53.25 85.31 18.70 1.25 52.8 0.5770.l27 3 Helical 0.375 144 57.00 1036 1455 5.971 54.11 90.86 19.96 1.3155.71 0.593 0.326 4 Helical 0.375 12 9.00 2792 3920 0.350 70.62 150.6024.41 1.79 85.34 0.745 0.911 4 Helical 0.375 12 9.00 3527 4952 0.27753.28 105.90 21.45 1.59 61.93 0.654 0.754 4 Helical 0.375 12 9.00 37035199 0.264 45.40 98.83 15.00 1.85 55.7 0.781 1.075 4 Helical 0.375 129.00 3487 4896 0.280 52.1l 113.70 17.34 1.85 64.37 0.789 1.150 4 Helical0.375 36 27.00 2888 4055 1.015 81.66 131.20 31.68 1.22 82.36 0.556 0.2265 Layered 0.375 5 2.50 1166 1637 0.233 109.60 248.20 29.14 2.00 133.50.769 0.682 5 Layered 0.375 5 2.50 2423 3402 0.112 29.37 58.21 6.71 1.7532.89 0.743 0.730 6 Helical 0.500 12 5.00 2764 2183 0.349 42.25 82.85l3.19 1.65 47.27 0.693 0.577 7 Helical 0.500 12 6.00 3929 3103 0.29521.51 43.89 4.75 1.82 24.22 0.731 0.546 7 Helical 0.500 12 6.00 35072770 0.330 29.67 57.92 7.59 1.70 32.68 0.705 0.499 7 Helical 0.500 126.00 2684 2120 0.431 40.03 71.89 12.50 1.48 42.05 0.638 0.283 7 Helical0.500 36 18.00 2779 2195 1.250 48.37 83.19 15.68 1.40 49.52 0.614 0.1898 Scissors 0.313 12 2.88 909 1838 0.238 23.66 48.68 4.88 1.85 26.750.742 0.579 8 Scissors 0.313 12 2.88 620 1254 0.350 58.77 104.90 20.111.44 62.83 0.650 0.574

The average emulsion velocity and residence time through a static mixercan be calculated from the flow rate into the static mixer and thelength and diameter of the static mixer as follows:

(1) Average Velocity=flow rate/cross-sectional area of empty staticmixer

(2) Residence Time=length of static mixer/Average Velocity

To characterize the particle size distribution, these parameters weremeasured: median, coefficient of variation of the median and skewness.The coefficient of variation of the median (CoV_(m)) was used tocharacterize the span of the distribution. $\begin{matrix}{{CoV}_{m} = \frac{( {{{diameter}\quad {of}\quad \mu \quad \theta \quad {at}\quad 84\%} - {{diameter}\quad {of}\quad \mu \quad \theta \quad {at}\quad 16\%}} )}{MMD}} & (3)\end{matrix}$

As the CoV_(m) approached zero, the distribution became moremonodispered. The symmetry of the distribution was determined using theskewness coefficient. The Coulter analyzer's method of determiningskewness is as follows:${Skewness} = \frac{\sum\lbrack {n_{c}( {x_{c} - x_{a}} )}^{3} )}{{SD}^{3}{\sum n_{c}}}$$\begin{matrix}{n_{c} = ~{\# \quad {of}\quad {particles}\quad {in}\quad {c'}{th}\quad {channel}}} \\{x_{c} = {{mean}\quad {volume}\quad {in}\quad {the}\quad {c'}{th}\quad {channel}}} \\{x_{a} = {{arithmetic}\quad {mean}\quad {of}\quad {the}\quad {distribution}}} \\{{SD} = {{standard}\quad {deviation}\quad {of}\quad {the}\quad {distribution}}}\end{matrix}$

As the distribution became more symmetrical, the skewness approachedzero. For right-skewed distributions (the particle size of the mode isless than the mean) the coefficient is positive; for left-skewed, thecoefficient is negative.

FIG. 4 shows the effect of the average emulsion velocity (cm/min) on themass median diameter (MMD, microns) of the size distribution for thevarious static mixers under test. As shown in FIG. 4, the MMD data forthe plastic helical design mixer indicate a consistent well behavedinverse relationship between MMD and average velocity, independent ofmixer diameter, length, and number of elements per length. The layeredstainless steel (SS) mixer and the plastic scissors mixer alsodemonstrated an inverse relationship between MMD and average velocity.FIG. 4 shows MMD data for a helical stainless steel (SS) mixer used toproduce microparticles at the one kilogram scale, in a manner analogousto that described above for the 100 gram scale. The inverse relationshipbetween MMD and average velocity was not readily apparent from theexperimental data obtained with the stainless steel helical mixer at theone kilogram scale. Notably, this static mixer had the fewest elementsper length (1.33 elements per inch) of all the static mixers tested.

FIG. 5 shows the effect of emulsion residence time (sec) in the staticmixer on the width of the microparticle size distribution (CoV_(m))using the various static mixers under test. An increase in residencetime of the emulsion flowing through the static mixer causes a decreasein the width of the microparticle size distribution, until a minimum isreached. The data at extremely low residence times (less than onesecond) have high variability, but consistently show an increase in thewidth of the particle size distribution as the residence time decreasesbelow one second.

FIG. 6 shows the effect of emulsion residence time (sec) in the staticmixer on the skewness of the microparticle size distribution. As shownin FIG. 6, as residence time increased, the skewness of themicroparticle size distribution decreased. All of the distributions wereright-skewed.

Experiments were also performed at the one kilogram scale, in a manneranalogous to that described above for the 100 gram scale. However, forthe one kilogram experiments, the number of static mixers was varied,using one, two, four, and eight static mixers arranged in series. Theresults of the one kilogram scale experiments are shown below in Table3.

TABLE 3 Flow Cal. Rate Cal. Residence 25-125 25-150 # of (ml/ VelocityTime Median @ 84% @ 16% @ 25 um @ 125 um @ 150 um yield* yield* Batch #Mixers min) (cm/min) (sec) (um) (um) (um) CoV_(m) % % % % % 1kg-0210-7 13060 4297 0.319 74.85 135.1 24.98 1.47 17.94 78.31 88.85 70.37 70.91-0218-7 1 3050 4283 0.320 79.95 135.4 29.85 1.32 N/R N/R N/R N/R N/R-0319-7 1 3050 4283 0.320 70.57 141 26.27 1.63 N/R N/R N/R N/R N/R-0325-7 1 3000 4212 0.326 71.51 122.3 24.95 1.36 14.96 77.11 85.88 62.1570.92 -0506-7 2 3560 4999 0.549 67.4 113.1 22.38 1.35 N/R N/R N/R N/RN/R -0708-7 4 3260 3325 1.650 59.38 97.22 24.99 1.22 N/R N/R N/R N/R N/R-0908-7 8 3140 3202 3.426 70.73 102.6 30.39 1.02 12.3 96.69 99.86 84.3987.56 -1014-7 8 3030 3090 3.551 80.57 121 30.37 1.12 13 86.3 95.96 73.382.96 Average 1 3040 4268 0.321 74.22 133.5 26.51 1.44 16.45 77.71 87.3761.26 70.92 Std. dev. 27 38 0.003 4.24 7.9 2.31 0.14   2.11  0.85  2.10 1.26  0.01 Average 8 3085 3146 3.489 75.65 111.80 30.38 1.07 12.6591.50 97.91 78.85 85.26 Std. dev. 78 79 0.088 6.96 13.01 0.01 0.07  0.49 7.35  2.76  7.84  3.25 *Theoretical yieid based on the particle sizeanalysis

Consistent with the 100 gram scale experiments, the results in Table 3show that the width of the distribution of the microparticle sizedecreases with increasing residence time of the emulsion in the seriesof static mixers. As shown in Table 3, the average residence time forone static mixer was 0.321 sec, resulting in a CoV_(m) of 1.44.Increasing the average residence time to 3.489 sec through the use ofeight mixers, decreased the CoV_(m) to 1.07. The longer averageresidence time also increased the average yield in a desiredmicroparticle size range of 25-125 μm by 28.71%, and in a desiredmicroparticle size range of 25-150 μm by 20.22%.

The experiments and data described above demonstrate that the massmedian diameter of the microparticles is inversely proportional to theaverage emulsion velocity through the static mixer. The experiments anddata also demonstrate that the width of the microparticle sizedistribution (CoV_(m)), and its symmetry, can be controlled by theresidence time of the emulsion in the static mixer, or series of staticmixers. Helical design static mixers, scissors design static mixers, andlayered design static mixers all appear to behave similarly with respectto changes in velocity or residence time.

Example 2 Preparation of Risperidone Microparticles Using a Manifold

Microparticles comprising risperidone were prepared at the 20-kilogramscale. The 20 Kg process (8 Kg of active agent and 12 Kg of polymer)provides a theoretical drug loading of the microparticles of 40% (8Kg/20 Kg×100%).

The polymer solution was prepared by dissolving 12.0 Kg of MEDISORB®7525 DL polymer (Alkermes, Inc., Blue Ash, Ohio) in 60 Kg of ethylacetate (Merck). The polymer was added to the solvent at 25° C. in astainless steel reactor. The temperature of the tank was raised to 37°C. to facilitate dissolution. The vessel was agitated for at least 16hours to dissolve the polymer. Once dissolved, the solution temperaturewas reduced to 25° C.

The drug solution was prepared by dissolving 8.0 Kg of risperidone base(Janssen Pharmaceutica, Beerse, Belgium) in 25.3 Kg of benzyl alcohol(Merck) at 25° C. in a stainless steel reactor. The organic phase wasprepared by adding the drug solution to the polymer solution at 25° C.in a stainless steel reactor and mixing for at least 15 minutes.

The continuous or aqueous phase was prepared by dissolving 6.0 Kg ofpolyvinyl alcohol (PVA) (DuPont) in 594.0 Kg hot (>60° C.) water forinjection (WFI) in a stainless steel vessel to form a 1% solution. Thevessel was agitated for at least 6 hours to dissolve the PVA. Once thePVA was dissolved, the temperature was reduced to 25° C. and 42.0 Kg ofethyl acetate (Merck) was added, and mixed for at least 30 minutes todissolve the ethyl acetate.

Using positive displacement pumps, the organic and aqueous phases werepumped through a static mixing assembly to form an oil-in-wateremulsion. A preferred system for carrying out this process is shown inFIG. 7. An organic phase 710 (polymer/drug solution) is pumped by a pump712. A continuous phase 720 (PVA solution) is pumped by a pump 722. Theorganic and continuous phases are pumped into a static mixing assembly730. Preferably, static mixing assembly 730 comprises a preblendingstatic mixer 732 and a manifold 734. Manifold 734 preferably comprises aplurality of individual static mixers 736 configured to provide aplurality of parallel flow streams 738. Although FIG. 7 shows twoparallel flow streams in manifold 734, the present invention is notlimited to the use of two flow streams, and more or less flow streamscould be used in manifold 734, as would be readily apparent to oneskilled in the art. Similarly, FIG. 7 shows three individual staticmixers 736 arranged in series to form flow streams 738. It should bereadily apparent to one skilled in the art that other numbers, more orless, of static mixers 736 could be used to form flow streams 738.

The emulsion droplet size, and the resulting microparticle size, arecontrolled primarily by the flow rate and residence time through thestatic mixing assembly. The static mixing assembly used in the 20 Kgrisperidone process consisted of a two inch diameter preblending staticmixer upstream of a two flow stream manifold. Each flow stream in themanifold was formed by three, eighteen inch long, one inch diameterstatic mixers arranged in series. The larger diameter of the preblendingstatic mixer is important to accommodate the total flow from the organicand aqueous phase feed streams at a lower velocity than the smallerdiameter static mixers in the manifold.

The flow rates are set by nominal pump speed settings. The total flowrate was approximately 20-22 Kg/min, divided between the two flowstreams in the manifold. The total flow rate can be divided one-half foreach flow stream in the manifold, or in any other manner as would bereadily apparent to one skilled in the art.

The flow rates for seven 20 Kg batches of microparticles are shown belowin Table 4. In batches 03098, 03168, and 03238, the drug (risperidone)was not present, resulting in the production of “placebo”microparticles.

TABLE 4 20 kg Emulsion flow rate summary w:o ratio Rate water Rate oilRate Total Batch w/w kg/min kg/min kg/min -03098 4.84 16.8 3.47 20.3-03168 4.84 16.8 3.47 20.3 -03238 4.86 16.8 3.47 20.3 -03308 4.70 16.83.57 20.3 -04068 4.67 17.0 3.63 20.6 -04138 4.50 16.9 3.75 20.7 -042084.46 16.8 3.78 20.6 Average 4.7 16.8 3.6 20.4 SD 0.2 0.1 0.1 0.2

After exiting the static mixing assembly, the oil-in-water emulsion istransferred to a large agitated tank for solvent extraction or quench(extraction vessel 740 in FIG. 7). The aqueous quench consisted of 4875Kg WFI and 125 Kg ethyl acetate at 6° C. Solvents are extracted from theemulsion droplets, thereby hardening into microparticles. Themicroparticles were sampled from the extraction vessel and analyzedusing a Coulter LQ100 particle size analyzer. The particle size resultsfrom these experiments are shown below in Table 5.

TABLE 5 Particle Size Characteristics Batch # Mean (um) Std. Dev. % <150 um 0309-8 83.03 40.44 95.7 0316-8 80.14 43.79 94.5 0323-8 80.8140.81 95.8 0330-8 82.24 36.9 97.7 0406-8 78.21 34.53 99 0413-8 80.4735.31 98.5 0420-8 79.55 35.75 98.5 Average 80.64 38.22 97.10 Std. Dev1.62 3.48 1.75

As can be seen from Table 5, the percentage of microparticles within thedesired microparticle size range of less than 150 μm ranged from 94.5%to 99%, with an average of 97.1%. Through the use of the static mixingassembly, the particle size characteristics could be controlled toprovide a desired microparticle size range.

It should be understood by one skilled in the art that the presentinvention is not limited to the use of an individual static mixer forany of the elements depicted as individual static mixers in the Figures.As would be readily apparent to one skilled in the art, a plurality ofindividual static mixers arranged in series could be used, or a manifoldcontaining a plurality of individual static mixers arranged in series,or arranged in parallel to provide a plurality of parallel flow streams,could also be used.

Example 3 Methods for Preparing Microparticles

As exemplified by the examples discussed above, methods for preparingmicroparticles in accordance with the present invention will now bedescribed in more detail. In one embodiment of the present invention, afirst phase, comprising an active agent and a polymer, is prepared. Inone embodiment of the present invention, the first phase is prepared bydissolving the active agent in a first solvent to form an active agentsolution. The polymer is dissolved in a second solvent to form a polymersolution. The active agent solution and the polymer solution are blendedto form the first phase. In a particularly preferred embodiment, theactive agent is selected from the group consisting of risperidone,9-hydroxyrisperidone, and pharmaceutically acceptable salts thereof. Insuch an embodiment, a preferred first solvent is benzyl alcohol, and apreferred second solvent is ethyl acetate.

In another embodiment of the present invention, the first phase isprepared by dissolving the active agent and the polymer in a solvent toform a solution. In a particularly preferred embodiment, the activeagent is bupivacaine, and the solvent is ethyl acetate. It should beunderstood that the present invention is not limited to any particularmethod or process by which the first phase is prepared, and othersuitable processes would be readily apparent to one skilled in the art.

A second phase is prepared, and combined with the first phase in a firststatic mixer to form an emulsion. In a preferred embodiment, the twophases are pumped into the static mixer, with the second phase beingpumped at a flow rate greater than the flow rate of the first phase. Inone preferred embodiment, the ratio of the flow rate of the second phaseto the flow rate of the first phase is from about 4:1 to about 5:1.However, it should be understood by one skilled in the art that thepresent invention is not limited to such a flow rate ratio, and otherappropriate flow rate ratios would be readily apparent to one skilled inthe art.

The emulsion flows through a manifold that includes a plurality ofstatic mixers into a quench liquid whereby droplets of the emulsion formmicroparticles. The quench liquid is prepared in a manner well known toone skilled in the art. The quench may be water, such as WFI or it maybe an aqueous solution comprising one or more solvents.

Preferably, the diameter of the first static mixer is greater than thediameter of each of the plurality of static mixers in the manifold. Inone embodiment, the plurality of static mixers is configured to providea plurality of parallel flow streams. In a particularly preferredembodiment, the plurality of static mixers in the manifold is two.However, it should be understood by one skilled in the art that thepresent invention is not limited to the use of two static mixers in sucha configuration, and other appropriate numbers of static mixers would bereadily apparent to one skilled in the art. Alternatively, the pluralityof static mixers can be configured in series to provide a plurality ofsequential flow streams.

The step of flowing the emulsion through the manifold is preferablyperformed by flowing a portion of a total flow rate through each of thestatic mixers in the manifold, the total flow rate being the sum of theflow rate of the first phase and the flow rate of the second phase. Forexample, for a manifold configured with two static mixers, a portion ofthe total flow rate will flow through each of the two static mixers. Theportion flowing through each of the two static mixers can be one-half,or other portions as would be readily apparent to one skilled in theart.

An alternate method for preparing microparticles in accordance with thepresent invention will now be described. A first phase, comprising anactive agent and a polymer, is prepared. A second phase is prepared, andcombined with the first phase in a first static mixer to form anemulsion, the emulsion forming an outflow of the first static mixer.Suitable methods and processes for preparing the first and secondphases, and for combining in the first static mixer, have been describedabove and will not be repeated here for brevity.

The outflow of the first static mixer is divided to form at least twoflow streams. Each of the at least two flow streams flows through aseparate second static mixer. The separate second static mixer can be anindividual static mixer, one of a plurality of individual static mixersarranged in series, or one of a plurality of individual static mixersconfigured to provide a plurality of parallel flow streams. Preferably,the diameter of the first static mixer is greater than the diameter ofeach separate second static mixer. The at least two flow streams mayhave substantially equal flow rates, or such other flow rates as wouldbe apparent to one skilled in the art.

The at least two flow streams are combined with a quench liquid wherebydroplets of the emulsion form microparticles. The quench liquid can bethe same as, or different from, the second phase.

A method for controlling particle size distribution of microparticles inaccordance with the present invention will now be described. In a manneras described above, a first phase, comprising an active agent and apolymer, is prepared. A second phase is prepared, and combined with thefirst phase in a static mixing assembly to form an emulsion. Theemulsion flows into a quench liquid whereby droplets of the emulsionform microparticles. The residence time of the emulsion in the staticmixing assembly is adjusted to obtain a predetermined particle sizedistribution of the resulting microparticles. The residence time isequal to a length of the static mixing assembly divided by an averagevelocity of the emulsion through the static mixing assembly. Increasingthe residence time of the emulsion in the static mixing assembly narrowsthe particle size distribution. Similarly, decreasing the residence timeof the emulsion in the static mixing assembly broadens the particle sizedistribution. The residence time may be adjusted by changing the lengthof the static mixing assembly. In one embodiment, the residence time isfrom about three seconds to about four seconds. However, it should beunderstood by one skilled in the art that the present invention is notlimited to such a residence time, and other residence times may be used.

In one embodiment of the present invention, the static mixing assemblycomprises a plurality of individual static mixers configured so that theemulsion flows sequentially through the plurality of individual staticmixers. In an alternate embodiment, the static mixing assembly comprisesa first static mixer and a manifold, in fluid communication with thefirst static mixer, which includes a plurality of static mixers. In suchan alternate embodiment, the diameter of the first static mixer ispreferably greater than the diameter of each of the plurality of staticmixers in the manifold.

Microparticles of the Present Invention

The microparticles prepared by the process of the present inventionpreferably comprise a polymeric binder, but it should be understood byone skilled in the art that the present invention is not limited topreparation of microparticles comprising a polymeric binder. Suitablepolymeric binder materials include poly(glycolic acid), poly-d,1-lacticacid, poly-1-lactic acid, copolymers of the foregoing, poly(aliphaticcarboxylic acids), copolyoxalates, polycaprolactone, polydioxanone,poly(ortho carbonates), poly(acetals), poly(lactic acid-caprolactone),polyorthoesters, poly(glycolic acid-caprolactone), polyanhydrides,polyphosphazines, albumin, casein, and waxes. Poly(d,1-lactic-co-glycolic acid) is commercially available from Alkermes,Inc. (Blue Ash, Ohio). A suitable product commercially available fromAlkermes, Inc. is a 50:50 poly(d,1-lactic-co-glycolic acid) known asMEDISORB® 5050 DL. This product has a mole percent composition of 50%lactide and 50% glycolide. Other suitable commercially availableproducts are MEDISORB® 6535 DL, 7525 DL, 8515 DL and poly(d,1-lacticacid) (100 DL). Poly(lactide-co-glycolides) are also commerciallyavailable from Boehringer Ingelheim (Germany) under its Resomer® mark,e.g., PLGA 50:50 (Resomer® RG 502), PLGA 75:25 (Resomer® RG 752) andd,1-PLA (Resomer® RG 206), and from Birmingham Polymers (Birmingham,Ala.). These copolymers are available in a wide range of molecularweights and ratios of lactic acid to glycolic acid.

One type of microparticle suitable for preparation by the presentinvention is a sustained-release microparticle that is biodegradable.However, it should be understood by one skilled in the art that thepresent invention is not limited to biodegradable or other types ofsustained-release microparticles. As would be apparent to one skilled inthe art, the molecular weight of the polymeric binder material forbiodegradable microparticles is of some importance. The molecular weightshould be high enough to permit the formation of satisfactory polymercoatings, i.e., the polymer should be a good film former. Usually, asatisfactory molecular weight is in the range of 5,000 to 500,000daltons, preferably about 150,000 daltons. However, since the propertiesof the film are also partially dependent on the particular polymericbinder material being used, it is very difficult to specify anappropriate molecular weight range for all polymers. The molecularweight of the polymer is also important from the point of view of itsinfluence upon the biodegradation rate of the polymer. For a diffusionalmechanism of drug release, the polymer should remain intact until all ofthe drug is released from the microparticles and then degrade. The drugcan also be released from the microparticles as the polymeric binderbioerodes. By an appropriate selection of polymeric materials amicroparticle formulation can be made in which the resultingmicroparticles exhibit both diffusional release and biodegradationrelease properties. This is useful in according multiphasic releasepatterns.

The microparticles prepared in accordance with the present invention mayinclude an active agent or other type of substance that is released fromthe microparticles into the host. Such active agents can include1,2-benzazoles, more particularly, 3-piperidinyl-substituted1,2-benzisoxazoles and 1,2-benzisothiazoles. The most preferred activeagents of this kind are3-[2-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]ethyl]-6,7,8,9-tetrahydro-2-methyl-4H--pyrido[1,2-a]pyrimidin-4-one(“risperidone”) and3-[2-[4-(6-fluro-1,2-benzisoxazol-3-yl)-1-piperidinyl]ethyl]-6,7,8,9-tetrahydro-9-hydroxy-2-methyl-4H--pyrido[1,2-a]pyrimidin-4-one(“9-hydroxyrisperidone”) and the pharmaceutically acceptable saltsthereof. Risperidone (which term, as used herein, is intended to includeits pharmaceutically acceptable salts) is most preferred. Risperidonecan be prepared in accordance with the teachings of U.S. Pat. No.4,804,663, the entirety of which is incorporated herein by reference.9-hydroxyrisperidone can be prepared in accordance with the teachings ofU.S. Pat. No. 5,158,952, the entirety of which is incorporated herein byreference.

Other biologically active agents include non-steroidal antifertilityagents; parasympathomimetic agents; psychotherapeutic agents; majortranquilizers such as chlorpromazine HCl, clozapine, mesoridazine,metiapine, reserpine, thioridazine and the like; minor tranquilizerssuch as chlordiazepoxide, diazepam meprobamate, temazepam and the like;rhinological decongestants; sedative-hypnotics such as codeine,phenobarbital, sodium pentobarbital, sodium secobarbital and the like;steroids such as testosterone and tesosterone propionate; sulfonamides;sympathomimetic agents; vaccines; vitamins and nutrients such as theessential amino acids; essential fats and the like; antimalarials such4-aminoquinolines, 8-aminoquinolines, pyrimethamine and the like,anti-migraine agents such as mazindol, phentermine and the like;anti-Parkinson agents such as L-dopa; anti-spasmodics such as atropine,methscopolamine bromide and the like; antispasmodics and anticholinergicagents such as bile therapy, digestants, enzymes and the like;antitussives such as dextromethorphan, noscapine and the like;bronchodilators; cardiovascular agents such as anti-hypertensivecompounds, Rauwolfia alkaloids, coronary vasodilators, nitroglycerin,organic nitrates, pentaerythritotetranitrate and the like; electrolytereplacements such as potassium chloride; ergotalkaloids such asergotamine with and without caffeine, hydrogenated ergot alkaloids,dihydroergocristine methanesulfate, dihydroergocornine methanesulfonate,dihydroergokroyptine methanesulfate and combinations thereof; alkaloidssuch as atropine sulfate, Belladonna, hyoscine hydrobromide and thelike; analgetics, narcotics such as codeine, dihydrocodienone,meperidine, morphine and the like; non-narcotics such as salicylates,aspirin, acetaminophen, d-propoxyphene and the like; antibiotics such assalicylates, aspirin, acetaminophen, d-propoxyphene and the like;antibiotics such as the cephalosporins, chloranphenical, gentamicin,Kanamycin A, Kanamycin B, the penicillins, ampicillin, streptomycin A,antimycin A, chloropamtheniol, metromidazole, oxytetracycline penicillinG, the tetracylines, and the like, anti-cancer agents; anti-convulsantssuch as mephenytoin, phenobarbital, trimethadione; anti-emetics such asthiethylperazine; antihistamines such as chlorophinazine,dimenhydrinate, diphenhydramine, perphenazine, tripelennamine and thelike; anti-inflammatory agents such as hormonal agents, hydrocortisone,prednisolone, prednisone, non-hormonal agents, allopurinol, aspirin,indomethacin, phenylbutazone and the like; prostaglandins; cytotoxicdrugs such as thiotepa; chlorambucil, cyclophosphamide, melphalan,nitrogen mustard, methotrexate and the like; antigens of suchmicroorganisms as Neisseria gonorrhea, Mycobacterium tuberculosis,Herpes virus (homonis, types 1 and 2), Candida albicans, Candidatropicalis, Trichomonas vaginalis, Haemophilus vaginalis, Group BStreptococcus ecoli, Mycoplasma hominis, Haemophilus ducreyi, Granulomainguinale, Lymphopathia venereum, Treponema pallidum, Brucella abortus,Brucella melitensis, Brucella suis, Brucella canis, Campylobacter fetus,Campylobacter fetus intestinalis, Leptospira pomona, Listeriamonocytogenes, Brucella ovis, Equine herpes virus 1, Equine arteritisvirus, IBR-IBP virus, BVD-MB virus, Chlamydia psittaci, Trichomonasfoetus, Toxoplasma gondii, Escherichia coli, Actinobacillus equuli,Salmonella abortus ovis, Salmonella abortus equi, Pseudomonasaeruginosa, Corynebacterium equi, Corynebacterium pyogenes,Actinobacillus seminis, Mycoplasma bovigenitalium, Aspergillusfumigatus, Absidia ramosa, Trypanosoma equiperdum, Babesia caballi,Clostridium tetani, and the like; antibodies that counteract the abovemicroorganisms; and enzymes such as ribonuclease, neuramidinase,trypsin, glycogen phosphorylase, sperm lactic dehydrogenase, spermhyaluronidase, adenosinetriphosphatase, alkaline phosphatase, alkalinephosphatase esterase, amino peptidase, trypsin, chymotrypsin, amylase,muramidase, acrosomal proteinase, diesterase, glutamic aciddehydrogenase, succinic acid dehydrogenase, beta-glycophosphatase,lipase, ATP-ase alpha-peptate gamma-glutamylotranspeptidase,sterol-3-beta-ol-dehydrogenase, and DPN-di-aprorasse.

Other suitable active agents include estrogens such as diethylstilbestrol, 17-beta-estradiol, estrone, ethinyl estradiol, mestranol,and the like; progestins such as norethindrone, norgestryl, ethynodioldiacetate, lynestrenol, medroxyprogesterone acetate, dimesthisterone,megestrol acetate, chlormadinone acetate, norgestimate, norethisterone,ethisterone, melengestrol, norethynodrel and the like; and thespermicidal compounds such as nonylphenoxypolyoxyethylene glycol,benzethonium chloride, chlorindanol and the like.

Still other suitable active agents include antifungals, antivirals,anticoagulants, anticonvulsants, antidepressants, antihistamines,hormones, vitamins and minerals, cardiovascular agents, peptides andproteins, nucleic acids, immunological agents, antigens of suchbacterial organisms as Streptococcus pneumoniae, Haemophilus influenzae,Staphylococcus aureus, Streptococcus pyogenes, Corynebacteriumdiphtheriae, Bacillus anthracis, Clostridium tetani, Clostridiumbotulinum, Clostridium perfringens, Streptococcus mutans, Salmonellatyphi, Haemophilus parainfluenzae, Bordetella pertussis, Francisellatularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila,Mycobacterium leprae, Leptospira interrogans, Borrelia burgdorferi,Campylobacter jejuni, antigens of such viruses as smallpox, influenza Aand B, respiratory syncytial, parainfluenza, measles, HIV,varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr,rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps,rabies, rubella, coxsackieviruses, equine encephalitis, Japaneseencephalitis, yellow fever, Rift Valley fever, lymphocyticchoriomeningitis, hepatitis B, antigens of such fungal protozoan, andparasitic organisms such as Cryptococcus neoformans, Histoplasmacapsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides,Rickettsia ricketsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydiapsittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosomabrucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis,Schistosoma mansoni. These antigens may be in the form of whole killedorganisms, peptides, proteins, glycoproteins, carbohydrates, orcombinations thereof.

Still other macromolecular bioactive agents that may be chosen forincorporation include, but are not limited to, blood clotting factors,hemopoietic factors, cytokines, interleukins, colony stimulatingfactors, growth factors, and analogs and fragments thereof.

The microparticles can be mixed by size or by type. However, it shouldbe understood that the present invention is not limited to the use ofbiodegradable or other types of microparticies that contain an activeagent. In one embodiment, the microparticles are mixed in a manner thatprovides for the delivery of active agent to the patient in amultiphasic manner and/or in a manner that provides different activeagents to the patient at different times, or a mixture of active agentsat the same time. For example, secondary antibiotics, vaccines, or anydesired active agent, either in microparticle form or in conventional,unencapsulated form can be blended with a primary active agent andprovided to the patient.

CONCLUSION

While various embodiments of the present invention have been describedabove, it should be understood that they have been presented by way ofexample only, and not limitation. The present invention is not limitedto the preparation of controlled release microparticles, nor is itlimited to a particular active agent, polymer or solvent, nor is thepresent invention limited to a particular scale or batch size. Thepresent invention is not limited to any type or design of static mixer.Thus, the breadth and scope of the present invention should not belimited by any of the above-described exemplary embodiments, but shouldbe defined only in accordance with the following claims and theirequivalents.

What is claimed is:
 1. A method of preparing microparticles, comprising:preparing a first phase, the first phase comprising an active agent anda polymer; preparing a second phase; preparing a quench liquid; pumpingthe first phase and the second phase through a first static mixer toform an emulsion; and flowing the emulsion through a manifold thatincludes a plurality of static mixers into the quench liquid wherebydroplets of the emulsion form microparticles.
 2. The method of claim 1,wherein a diameter of the first static mixer is greater than a diameterof each of the plurality of static mixers in the manifold.
 3. The methodof claim 1, wherein the pumping step is performed wherein the firstphase is pumped at a first flow rate and the second phase is pumped at asecond flow rate greater than the first flow rate.
 4. The method ofclaim 3, wherein a ratio of the second flow rate to the first flow rateis from about 4:1 to about 5:1.
 5. The method of claim 3, wherein theflowing step is performed by flowing a portion of a total flow ratethrough each of the plurality of static mixers in the manifold, whereinthe total flow rate is the sum of the first flow rate and the secondflow rate.
 6. The method of claim 5, wherein the plurality of staticmixers in the manifold is two.
 7. The method of claim 1, wherein thefirst static mixer comprises a plurality of static mixing elementsreceived within a conduit.
 8. The method of claim 7, wherein each of theplurality of static mixers in the manifold comprises a plurality ofstatic mixing elements received within a conduit.
 9. The method of claim1, wherein the step of preparing the first phase comprises: dissolvingthe active agent in a first solvent to form an active agent solution;dissolving the polymer in a second solvent to form a polymer solution;and blending the active agent solution and the polymer solution.
 10. Themethod of claim 9, wherein the active agent is selected from the groupconsisting of risperidone, 9-hydroxyrisperidone, and pharmaceuticallyacceptable salts thereof.
 11. The method of claim 10, wherein the firstsolvent is benzyl alcohol.
 12. The method of claim 10, wherein thepolymer is poly(d,l-lactide-co-glycolide) having a molar ratio oflactide to glycolide in the range of from about 85:15 to about 50:50.13. The method of claim 12, wherein the second solvent is ethyl acetate.14. A method of preparing microparticles, comprising: preparing a firstphase, the first phase comprising an active agent and a polymer;preparing a second phase; preparing a quench liquid; combining the firstphase and the second phase in a first static mixer to form an emulsion,the emulsion forming an outflow of the first static mixer; dividing theoutflow of the first static mixer to form at least two flow streams;flowing each of the at least two flow streams through a separate secondstatic mixer; and combining the at least two flow streams with thequench liquid whereby droplets of the emulsion form microparticles. 15.The method of claim 14, wherein a diameter of the first static mixer isgreater than a diameter of each separate second static mixer.
 16. Themethod of claim 14, wherein the step of preparing the first phasecomprises: dissolving the active agent in a first solvent to form anactive agent solution; dissolving the polymer in a second solvent toform a polymer solution; and blending the active agent solution and thepolymer solution.
 17. The method of claim 16, wherein the active agentis selected from the group consisting of risperidone,9-hydroxyrisperidone, and pharmaceutically acceptable salts thereof. 18.The method of claim 17, wherein the first solvent is benzyl alcohol. 19.The method of claim 16, wherein the polymer ispoly(d,l-lactide-co-glycolide) having a molar ratio of lactide toglycolide in the range of from about 85:15 to about 50:50.
 20. Themethod of claim 19, wherein the second solvent is ethyl acetate.
 21. Themethod of claim 14, wherein the at least two flow streams havesubstantially equal flow rates.
 22. A microencapsulated active agentprepared by a method for preparing microparticles, the methodcomprising: preparing a first phase, the first phase comprising anactive agent and a polymer; preparing a second phase; preparing a quenchliquid; pumping the first phase and the second phase through a firststatic mixer to form an emulsion; and flowing the emulsion through amanifold that includes a plurality of static mixers into the quenchliquid whereby droplets of the emulsion form microparticles.
 23. Amicroencapsulated active agent prepared by a method for preparingmicroparticles, the method comprising: preparing a first phase, thefirst phase comprising an active agent and a polymer; preparing a secondphase; preparing a quench liquid; combining the first phase and thesecond phase in a first static mixer to form an emulsion, the emulsionforming an outflow of the first static mixer; dividing the outflow ofthe first static mixer to form at least two substantially identical flowstreams; flowing each of the at least two substantially identical flowstreams through a separate second static mixer; and combining the leasttwo substantially identical flow streams with the quench liquid wherebydroplets of the emulsion form microparticles.
 24. A system for preparingmicroparticles, comprising: a first pump; a second pump; a first staticmixer in fluid communication with said first pump and with said secondpump, wherein said first pump is configured to pump an organic phaseinto said first static mixer, and said second pump is configured to pumpa continuous phase into said first static mixer; a manifold in fluidcommunication with said first static mixer, said manifold comprising aplurality of static mixers; and an extraction vessel in fluidcommunication with said manifold, wherein an outflow of said firststatic mixer flows through said manifold into said extraction vessel.25. The system of claim 24, wherein said first pump is configured tooperate at a first flow rate, and said second pump is configured tooperate at a second flow rate greater than said first flow rate.
 26. Thesystem of claim 25, wherein a ratio of said second flow rate to saidfirst flow rate is from about 4:1 to about 5:1.
 27. The system of claim24, wherein a diameter of said first static mixer is greater than adiameter of each of said plurality of static mixers in said manifold.28. The system of claim 24, wherein said plurality of static mixers insaid manifold is two.
 29. The system of claim 27, wherein said pluralityof static mixers in said manifold is two.
 30. The system of claim 24,further comprising: a first vessel in fluid communication with saidfirst pump; and a second vessel in fluid communication with said secondpump.
 31. A method for controlling particle size distribution ofmicroparticles, comprising: preparing a first phase, the first phasecomprising an active agent and a polymer; preparing a second phase;preparing a quench liquid; pumping the first phase and the second phasethrough a static mixing assembly to form an emulsion; flowing theemulsion into the quench liquid whereby droplets of the emulsion formmicroparticles; and adjusting a residence time of the emulsion in thestatic mixing assembly to obtain a predetermined particle sizedistribution of the resulting microparticles, wherein the residence timeis equal to a length of the static mixing assembly divided by an averagevelocity of the emulsion through the static mixing assembly.
 32. Themethod of claim 31, wherein the adjusting step is carried out toincrease the residence time, thereby narrowing particle sizedistribution.
 33. The method of claim 31, wherein the adjusting step iscarried out to decrease the residence time, thereby broadening particlesize distribution.
 34. The method of claim 31, wherein the static mixingassembly comprises a plurality of individual static mixers configured sothat the emulsion flows sequentially through the plurality of individualstatic mixers.
 35. The method of claim 31, wherein the static mixingassembly comprises: a first static mixer; and a manifold that includes aplurality of static mixers, the manifold in fluid communication with thefirst static mixer.
 36. The method of claim 35, wherein a diameter ofthe first static mixer is greater than a diameter of each of theplurality of static mixers in the manifold.
 37. The method of claim 31,wherein the residence time is from about three seconds to about fourseconds.
 38. The method of claim 35, wherein the residence time is fromabout three seconds to about four seconds.
 39. The method of claim 31,wherein the adjusting step is carried out by changing the length of thestatic mixing assembly.
 40. The method of claim 6, wherein the portionof the total flow rate flowing through each of the two static mixers isone-half.
 41. The method of claim 35, wherein the plurality of staticmixers in the manifold are configured to provide a plurality of parallelflow streams.
 42. The method of claim 36, wherein the plurality ofstatic mixers in the manifold are configured to provide a plurality ofparallel flow streams.
 43. The system of claim 24, wherein saidplurality of static mixers are configured in parallel.
 44. The system ofclaim 24, wherein said plurality of static mixers are configured inseries.